Malignant melanoma which belongs to a highly malignant tumor derived from melanocytes, is a common malignant tumor in skin cancer and has a strong metastasis. The main treatment method is radiotherapy, chemotherapy and surgical treatment, however, such traditional treatments cannot completely cure it. With the development of immunology and molecular biology, the tumor immunotherapy has become a hot topic of research. Toll like receptor (TLRs), an important class of pattern recognition receptors, belongs to type I transmembrane proteins, and is expressed in various immune cells. TLRs play an important role in stimulating the process of innate and adaptive immune responses. The agonists of TLRs act on antigen presenting cells, such as dendritic Cell (DC), to promote the production of cytokines and thus better stimulate the immune response. TLR7 (Toll like receptor 7) is one of the Toll like receptors whose ligand is mainly a nucleic acid component of the virus, and it can also identify some synthetic small molecular agonists. A stronger immune response can be triggered when TLR7 is activated. T7 (the first reagent) is an agonist of a small chemical molecule TLR7 receptor synthesized by the Shenzhen synthetic biology engineering laboratory of Shenzhen University. It is reported that T7 has the ability to improve innate immunity. When it is reacted with dendritic cells derived from bone marrow cells in vitro, inflammatory mediators, such as TNF-a and IL-12 can be produced rapidly, meanwhile when it is reacted with lymphocyte, inflammatory mediators, such as IFN-γ and IL-12 can also be produced rapidly. The source and preparation process of T7 may refers to “Local administration of a novel Toll-like receptor 7 agonist in combination with doxorubicin induces durable tumoricidal effects in a murine model of T cell lymphoma” Journal of Hematology & Oncology. 2015, Mar. 4; 8(1):21, doi: 10.1186/s13045-015-0121-9”. Ethacrynic acid (EA, the second reagent), also called as diuretic acid, is a diuretic. It has been reported that EA has a variety of anti-tumor effects, but it cannot stimulate the specific anti-tumor cell immune response, which makes its anti-tumor effect poor and its side effects obvious, thus the tumor cells relapse and grow repeatedly.
Mammary cancer is a malignant tumor occurring in the mammary gland epithelial tissue. The majority of the patients are female, and patients of advanced stage are prone to have tumor cells metastases. At present, the main treatments of mammary cancer are surgical treatment, radiotherapy and chemotherapy. However, the radiotherapy and chemotherapy can kill normal cells while killing tumor cells, which would seriously destroy the immune system of the human body. Tumor biotherapy is the fourth way of tumor treatment, in which the tumor immunotherapy is a promising and better treatment for mammary cancer. The purpose of tumor immunotherapy is to stimulate or mobilize the immune system of the human body, enhance the anti-tumor immunity of the tumor microenvironment, and induce the human body to produce immune cells that can effectively identify the tumor related antigen, so as to control and kill the tumor cells. ROR1 (Receptor-tyrosine-kinase-like orphan receptor, the third reagent) is a typical type I receptor-tyrosine-kinase-like orphan receptor surface protein. It is found that ROR1 is expressed in many human-derived tumor cell lines. Moreover, ROR1 is highly expressed in mammary tumor cells, but not expressed in the normal mammary cells, and the expression of ROR1 is related to the growth of the tumor cell. The higher the expression of ROR1, the faster the growth of the tumor. It is reported that the expression of ROR1 is closely related to the early metastasis, recurrence and prognosis of the mammary tumor. The patients with high expression of ROR1 have a short survival time without metastasis than those with low expression. Blocking the expression of ROR1 can significantly inhibit the metastasis of tumor cells to the lung. Therefore, ROR1 can be used to as a target for tumor therapy. However, ROR1 has a weak immunogenicity, so cannot stimulate innate immunity.